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*Compound via MeSH
*Substance via MeSH
Hazardous Substances DB
*12-O-TETRADECANOYLPHORBOL-13-ACETATE
*AMPHETAMINE
*CALCIUM COMPOUNDS
*CALCIUM, ELEMENTAL
*DOPAMINE

Vol. 297, Issue 3, 1016-1024, June 2001

Protein Kinase C and Intracellular Calcium Are Required for Amphetamine-Mediated Dopamine Release via the Norepinephrine Transporter in Undifferentiated PC12 Cells

Lana Kantor, G. H. Keikilani Hewlett, Yang Hae Park, Sarah M. Richardson-Burns, Mathew J. Mellon and Margaret E. Gnegy

Department of Pharmacology, University of Michigan School of Medicine, Ann Arbor, Michigan

The role of protein kinase C and intracellular Ca2+ on amphetamine-mediated dopamine release through the norepinephrine plasmalemmal transporter in undifferentiated PC12 cells was investigated. The selective protein kinase C inhibitor chelerythrine completely inhibited endogenous dopamine release elicited by 1 µM amphetamine. Direct activation of protein kinase C increased dopamine release in a Ca2+-insensitive, imipramine-sensitive manner and the release was not additive with amphetamine. Exocytosis was not involved since these events were not altered by either deletion of extracellular Ca2+ or reserpine pretreatment. Down-regulation of protein kinase C activity by long-term phorbol ester treatment resulted in a dramatic decrease in amphetamine-mediated dopamine release with no apparent effect on [3H]dopamine uptake. To more completely examine a role for Ca2+, intracellular Ca2+ was chelated in the cells. Depletion of intracellular Ca2+ considerably decreased dopamine release in response to 1 µM amphetamine compared with vehicle-treated cells, but had no effect on the [3H]dopamine uptake. Thus, our results suggest that amphetamine-mediated dopamine release through the plasmalemmal norepinephrine transporter is highly dependent on protein kinase C activity and intracellular but not extracellular Ca2+. Furthermore, protein kinase C and intracellular Ca2+ appear to regulate [3H]dopamine inward transport and amphetamine-mediated outward transport of dopamine independently in PC12 cells.

0022-3565/01/2973-1016$03.00/0
THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS
Copyright © 2001 by The American Society for Pharmacology and Experimental Therapeutics


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